Development of the UValidate platform for the profiling of topically applied chemical agents.

开发 UValidate 平台，用于分析局部应用的化学制剂。

• 批准号: 10484288
• 负责人: DEAN ROSENTHAL
• 金额: $ 80.92万
• 依托单位: AMELIA TECHNOLOGIES, LLC
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-20 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10484288
• 关键词: 3-Dimensional; Accounting; Address; Aging; Algorithms; Animals; Behavioral; Biological; Biological Assay; Biology; Blood; CRISPR/Cas technology; Catalogs; Cell Line; Cell model; Cells; Chemical Agents; Chemicals; Clustered Regularly Interspaced Short Palindromic Repeats; Complement; Complex; Computer software; Contract Services; Cosmetics; Country; Coupled; Cultured Cells; Cutaneous; DNA Damage; DNA Repair; DNA lesion; Data Storage and Retrieval; Dermal; Development; Diffusion; Digestion; Effectiveness; Ensure; Environment; Exposure to; FDA approved; Face; Fibroblasts; Formulation; Funding; Future; Health; Health Knowledge Attitudes Practice; Human; Individual; Industry; Kinetics; Knock-in; Label; Lead; Lesion; Malignant Neoplasms; Measurement; Measures; Minerals; Modeling; Molecular; Mutagenicity Tests; Mutation; Organ; Patients; Pharmaceutical Preparations; Phase; Poison; Population; Protective Clothing; Quality Control; Reactive Oxygen Species; Reagent; Reproducibility; Research Personnel; Risk; Safety; Services; Short Waves; Skin; Skin Aging; Skin Cancer; Skin Care; Skin tanning; Small Business Technology Transfer Research; Spectrometry; Sunscreening Agents; Surface; Technology; Testing; The Sun; Time; Topical agent; Topical application; UVA induced; Ultraviolet B Radiation; United States; Work; Zinc Oxide; alkalinity; base; cancer initiation; carcinogenesis; carcinogenicity; cell type; combinatorial; data acquisition; data pipeline; deep learning; design; drug discovery; high throughput screening; improved; individual response; irradiation; keratinocyte; melanocyte; novel; prevent; repaired; research clinical testing; screening; skin barrier; skin disorder; solar ultraviolet radiation; success; sun damage; three dimensional cell culture; titanium dioxide; ultraviolet irradiation

Project Summary Skin is the first line of defense against the outside environment. When the skin barrier is breached by UV irradiation (UVR) or toxic chemicals, living cell layers including epidermal keratinocytes, melanocytes and dermal fibroblasts are subjected to DNA damage. If unrepaired, this damage can lead to photoaging and cutaneous carcinogenesis. In fact, skin cancer is by far the most common malignancy, accounting for more cases of cancer in the US and other countries than all other organs combined. Sunscreens have been used extensively with some success, but newer chemical filters suffer from serious shortcomings that make currently available products undesirable, and can be a risk to human health. Current strategies that examine the effectiveness and safety of organic UV filters need to be reassessed. These new strategies should test the biology of UVR in the presence of sunscreens. Further, they need to be high-throughput (HTP) in order to examine combinations of different chemical blockers that are present in most formulations, work in cultured cells for ease of screening, and be animal-friendly. In Phase I we showed proof-of-principle for a novel UVR HTP screening platform called “UValidate, employing mixed populations of isogenic keratinocytes and melanocytes derived from a single donor, exposed to two-compound combinations of sunscreen active ingredients in the presence and absence of short wave (UVB; 285-320), and long wave solar UVR (UVA1; 340-400 nm). These screens measure reactive oxygen species (ROS), DNA damage (CPDs), and micronuclei and show a complex interplay between solar UV wavelength, sunscreen blockers, and cell types. This highlights the pressing need for our platform, compared to currently-utilized assays that measure only the amount of UV that is blocked by spectrometry. In the proposed Phase II, we plan to test known chemical UV blockers for the cosmetic and skin-care markets as a contracting service, using a panel of donor-derived isogenic keratinocytes, melanocytes and fibroblast. At the completion of Phase II, if funded, UValidate will be offered as a service particularly suited for companies at the early stages of drug discovery, regulatory bodies and established pharma seeking to optimize formulation. Advances in this proposal include fluoro-tagging isogenic patient cell lines, establishing 2D and 3D cell cultures to determine individual responses to solar UV irradiation, utilization of reagents and assays to rapidly determine the types of DNA lesions and their repair, and design of AI software to more accurately determine damage. Control cells, generated using CRISPR technology, will reproduce DNA repair-compromised skin cells and 3D skin equivalents will also be included to ensure experimental rigor and reproducibility. The present crisis due to the lack of non- toxic sunscreens has led to class action lawsuits, banning of sunscreens in growing numbers worldwide, and recall of formulations from the market. Therefore, this technology is timely and needed with great expediency to address the urgent need for the discovery and testing of safe and effective UV blockers to prevent the most common, and often debilitating and deadly cancers that we face.

项目摘要 皮肤是抵御外界环境的第一道防线。当皮肤屏障被紫外线破坏时 辐射（UVR）或有毒化学物质，活细胞层，包括表皮角质形成细胞、黑素细胞和真皮 成纤维细胞受到DNA损伤。如果不修复，这种损伤可能导致光老化和皮肤损伤。 致癌作用事实上，皮肤癌是迄今为止最常见的恶性肿瘤，占癌症病例更多 在美国和其他国家比所有其他器官的总和。防晒霜已广泛用于 一些成功，但较新的化学过滤器遭受严重的缺点，使目前可用的 产品不受欢迎，并且可能对人类健康构成风险。目前的战略，审查有效性和 有机紫外线过滤剂的安全性需要重新评估。这些新的策略应该测试紫外线辐射的生物学， 防晒霜的存在。此外，它们需要是高通量的（HTP），以便检查以下的组合： 存在于大多数制剂中的不同化学阻断剂，在培养的细胞中起作用以便于筛选， 善待动物在第一阶段，我们展示了一种新的UVR HTP筛选平台的原理验证， “Uceptide，采用来自单个供体的同基因角质形成细胞和黑素细胞的混合群体， 暴露于防晒活性成分的两种化合物组合， 长波紫外线（UVB; 285-320）和长波太阳紫外线（UVA 1; 340-400 nm）。这些屏幕测量活性氧 物种（ROS），DNA损伤（CPD）和微核，并显示太阳紫外线之间的复杂相互作用 波长、防晒剂和细胞类型。这凸显了对我们平台的迫切需求，相比之下， 到目前使用的仅测量被光谱法阻挡的UV的量的测定。拟议 第二阶段，我们计划测试已知的化学紫外线阻断剂的化妆品和皮肤护理市场，作为一个合同， 服务，使用一组供体来源的等基因角质形成细胞，黑素细胞和成纤维细胞。完成时 第二阶段，如果得到资助，UCNET将作为一项服务提供，特别适合处于早期阶段的公司， 药物发现、监管机构和寻求优化配方的老牌制药公司。这方面的进展 建议包括荧光标记同基因患者细胞系，建立2D和3D细胞培养物以确定 个人对太阳紫外线照射的反应，利用试剂和测定法快速确定 DNA损伤及其修复，以及设计AI软件以更准确地确定损伤。对照细胞， 使用CRISPR技术产生的，将复制DNA修复受损的皮肤细胞和3D皮肤等效物 也将包括在内，以确保实验的严谨性和可重复性。目前的危机是由于缺乏非 有毒的防晒霜导致了集体诉讼，在世界范围内越来越多的防晒霜被禁止， 从市场上召回配方。因此，这项技术是及时的，需要极大的便利， 满足发现和测试安全有效的紫外线阻断剂的迫切需要，以防止最严重的 常见的，通常是致命的癌症。